BIOLOGICAL RESOURCES UNITThe <strong>Cleveland</strong> <strong>Clinic</strong> Foundation’s Biological Resources Unit (BRU; formerly Animal CoreFacilities) is an integral unit of the <strong>Lerner</strong> <strong>Research</strong> Institute. The BRU’s function is to give support inmatters related to animal experimentation to all the scientists of the Foundation who use animals intheir research projects. This function is carried out with full consideration of the animals’ welfare. Thus,all efforts are made to minimize the number of animals used and any physical and psychological distressexperienced.The facilities are located in multiple areas of the CCF, but centrally managed. A variety of animalspecies, from rodents to ruminants, are housed in those areas. State-of-the-art surgical facilities andequipment are provided in each of the sites with cineangiography units (with digitizing and freeze-framecapabilities) and general instrumentation support. Approximately 30 animal technicians staff the BRU,many of them certified by the American Association for Laboratory Animal Science (AALAS).The services provided by the BRU encompass all aspects related to animal husbandry andexperimental manipulation, ordering and procurement of animals, quarantine procedures, perioperativeanimal care, health surveillance and consultation on animals models. Implementation of, and compliancewith, the Animal Welfare Act and Public Health Services policies are conducted in conjunction with theInstitutional Animal Care and Use Committee (IACUC). In 2001, the CCF animal care and use programwas evaluated by the Association for the Assessment and Accreditation of Laboratory Animal Care,International (AAALAC) and received full accreditation as part of an ongoing program. The BRU’sfacilities have been AAALAC accredited since 1976. During the past year, the BRU has implemented anew animal users’ training and occupational health program to meet NIH initiatives. Implementation ofincreased security measures is under way.The BRU will move into its new vivarium in the fall of <strong>2003</strong>. This facility will supplementexisting animal housing and allow for expansion of both the animal census and for services provided bythe BRU for all CCG investigators. It will also consolidate satellite housing areas that have providedlimited temporary expansion space.Website: http://info.lerner.ccf.org/services/bru/BIOLOGICAL RESOURCES UNITDIRECTORStanley D. Dannemiller, D.V.M., M.S., Dipl. A.C.L.A.M.ASSISTANT DIRECTORJori Leszczynski, D.V.M.182MANAGERSLinda McCort, A.S., L.A.T.G.Lonnie Thomas, B.A., L.A.T.G.RESEARCH COMPLIANCECOORDINATORNatalie L. Mays, B.A., L.A.T.G.CHIEF TECHNICIANHoward Roper, L.A.T., I.L.A.M.ANIMAL SURGERY SUPERVISORRobert LewisANIMAL SURGERY TECHNICIANSJames Howard, L.A.T.Terry Keller, L.A.T.Ronald Porter, L.A.T.MICHELLE WALSHCHRONIC CARE UNITTerry Fye (<strong>Research</strong> Monitor)VETERINARY TECHNICIANMelanie Hoffner, R.V.T.ANIMAL TECHNICIAN GROUP LEADERSJohn Bolden, A.L.A.T.China Morrison-KeetonScott Zeigler, L.A.T.ADMINISTRATIVE SUPPORTPeggy Buckner (Dept. Coordinator)Deborah Underwood (ClericalAsst.)ANIMAL TECHNICIANSPatrina BarlowDavid BredaDawn CadeJanie CallowayMary ColemanLaToya DavisPaul DevineStephanie EncarnacionDeborah FlorioJanice FranklinCharles FullerElizabeth HarmonDonnie HayesYvonne JamesYegeniy Kononov, L.A.T.Larry Kramer, Jr.Robert McCutcheonDrazen Mikulec, L.A.T.Mary MitchellRebecca ShrefflerJose SolanoDeAndre TerrellHenry Wyant
MOLECULAR BIOTECHNOLOGY COREThe Molecular Biotechnology Core laboratory provides consultation and technical support servicesin DNA sequencing protein analysis, peptide chemistry, amino acid analysis, and bioconjugate chemistry.These services are made available to all investigators of the <strong>Lerner</strong> <strong>Research</strong> Institute and the <strong>Cleveland</strong><strong>Clinic</strong> Foundation.High-throughput DNA sequencing systems based on capillary electrophoresis have revolutionizedthe genome sequencing projects dramatically in the last several years. The blueprints of human,Arabidopsis, Caenorhabditis elegans, Drosophila, yeast, Escherichia coli, Haemophilus influenzae, and severalother microbial genomes have now been sequenced. Similarly, the huge data-processing power ofcomputers and increasing availability of powerful laboratory data management software in recent yearshas made annotation of genome sequences possible in databases. This has stimulated mining of databases,using powerful technologies such as high-throughput mapping of single-nucleotide polymorphisms(SNPs) through the entire genome. SNPs are expected to provide clues to predisposition to disease andresponse to drugs. As a consequence, life scientists are using DNA sequencing as a routine researchpractice in their laboratories to understand fundamental biological processes at molecular level. Therefore,DNA sequencing will continue to be a major task in the foreseeable future as gene sequences aresurveyed more often. In the post genome era, the Molecular Biotechnology Core laboratory at the <strong>Lerner</strong><strong>Research</strong> Institute will continue to play an important role by providing crucial research support in theareas of customized DNA sequencing and SNP identification. The DNA sequencing samples submittedby CCF investigators to the Core has increased steadily. To meet this demand, a MegaBACE 1000workstation, which uses capillary array electrophoresis, became operational in January 2001.The core laboratory also offers N-terminal amino acid sequence analysis by Edman degradationchemistry and amino acid composition analysis by the orthophthalaldehyde (OPA) method. Facilities forin-gel digestion and peptide separation (mapping) of N-terminal blocked protein for internal peptidesequence analysis are available, although mass spectrometry is lately becoming a method of choice inprotein identification. In the area of peptide chemistry, the laboratory performs peptide synthesis by thesolid-phase method using Fmoc chemistry. To exploit the full potential of synthetic peptides in biomedicalresearch, the Core laboratory has an ABI 431A synthesizer (Applied Biosystems) and an Omega 396multiple peptide synthesizer (Advanced ChemTech). With the installation of the new 396 Omegamultiple peptide synthesizer, CCF investigators have the flexibility of synthesis at the 50-, 100- or 250-mmole scale. Every peptide synthesized is rigorously evaluated routinely by mass spectrometry, HPLCanalysis and by N-terminal sequencing if necessary with the ultimate goal of providing the correctintended peptide sequence to the investigator.MOLECULARBIOTECHNOLOGYCOREDIRECTORSatya Yadav, Ph.D.SCIENTIFIC ADVISOREDWARD PLOW, PH.D.SUPPORT PERSONNELTalat Haqqi, M.S.Adriana Panciu, B.S.Wei-Zhen Shen, B.S.Xiaolan Zhao, M.D.Web site: http://www.lerner.ccf.org/services/molecbiotech/VIRUS COREThe Virus Core Facility provides services and expertise, including scientific consultations, relatedto infectious and noninfectious viruses. The Core provides such essential services as making virus stocks,inducing large-scale expression of cloned proteins in baculovirus, constructing retroviral and adenoviralvectors expressing cDNAs, and making respective recombinant virus stocks.Currently, the core maintains the following virus stocks: Human immunodeficiency virus (HIV-1), respiratory syncytial virus (RSV), parainfluenza virus (PIV), recombinant baculovirus, adenovirus,retrovirus and other viruses as requested. For protein expression in baculovirus, primary services includeconstructing recombinant baculovirus expressing cloned cDNA (with or without His-tag), makingrecombinant virus and performing small-scale infections to characterize gene expression, determining theoptimum conditions of harvest time and infection ratio that gives maximum protein yield, and carryingout large-scale infection to produce large quantities of protein. The Core also offers a complete range ofservices for the construction of recombinant adenoviruses, including cloning, plaque purification,screening, amplification, double cesium chloride gradient purification, and viral particle titration.The Virus Core Facility is involved in new collaborative projects that will generate valuableresearch materials for the research institute. The Virus Core has trained numerous scientists in how togenerate recombinant baculoviruses, adenoviruses and retroviruses and has designed experiments fortheir research work. We are also in the process of making new adenoviral vectors that will regulate geneexpression in a tissue-specific manner.VIRUS COREDIRECTORRatan K. Maitra, Ph.D.SCIENTIFIC CONSULTANTSAmiya K. Banerjee, Ph.D.Joseph DiDonato, Ph.D.SUPPORT PERSONNELDonald R. RempinskiWeb site: http://www.lerner.ccf.org/services/virus/183